Effects of magnesium-substituted nanohydroxyapatite coating on implant osseointegration.

OBJECTIVE: The objective of this study was to compare magnesium-substituted and pure hydroxyapatite coatings on the promotion of osteogenesis in vitro and on the osseointegration in vivo. METHODS: Electrochemically deposited pure hydroxyapatite (EDHA) or electrochemically deposited magnesium-substituted hydroxyapatite (EDMHA) coatings were formed on the surface of pure titanium disks or implants. MC3T3-E1 preosteoblasts were cultured in the EDHA and EDMHA coated disks, and cell growth, alkaline phosphatase (ALP) activity, and osteocalcin secretion were measured at various time points. For studies on osseointegration, 30 roughened implants coated either with EDHA or EDMHA (n = 15 for each coating) were implanted in the femurs of 15 NZW rabbits. After 2, 4, and 8 weeks, femurs were retrieved and prepared for histomorphometric evaluation (n = 5 for each coating at each time point). RESULTS: MC3T3-E1 cells cultured on EDMHA coated disks showed increased cell number, ALP, and osteocalcin secretion compared with the EDHA coated disks at all time points (P < 0.05 for all). Histologic observation of the coated implants showed woven bone in direct contact with both implant surfaces after 2 weeks and mature bone after 8 weeks. While there were no differences in the amount of bone between the threads at any time point, the percentage of implant in direct contact with bone (bone implant contact) was slightly higher along the EDMHA coated implants at 2 weeks (P = 0.086), although this difference was no longer seen at 4 and 8 weeks. CONCLUSION: Mg-substituted HA coated surfaces promote osteogenic differentiation of preosteoblasts in vitro and may improve implant osseointegration during the early stages of bone healing compared with pure EDHA coated surfaces.

Bone response to the multilayer BMP-2 gene coated porous titanium implant surface.

OBJECTIVES: Evaluate hBMP-2 expression following gene delivery from plasmid multilayers formed on sandblasted titanium in vitro and bone formation around similarly prepared implant surfaces in vivo. MATERIALS AND METHODS: Multilayers of cationic lipid/rhBMP-2 plasmid DNA complex (LDc) and anionic hyaluronic acid (HA) was assembled on sandblasted-dual acid etched pure titanium disks or implant surfaces using layer-by-layer (LBL) assembly. Gene delivery and hBMP-2 expression in cells exposed to the LDc multilayers was measured in vitro. To determine the effect of BMP delivery from such multilyaers in vivo, roughened implants coated with BMP-2 LDc multilayers or uncoated control implants (n = 15 for both) were implanted in the femurs of NZW rabbits. After 2, 4, 8 weeks, femurs were retrieved and prepared for histomorphometric evaluation (n = 5 rabbits per time point). RESULTS: MC3T3-E1 cells cultured directly on the BMP-2 LDc coated titanium disks showed EGFP and hBMP-2 expression after 48 h in culture. Increased gene delivery occurred by increasing the number of assembly layers when cells were cultured for 48 h. Cells cultured on LDc coated surfaces had significantly higher cell viability than control cells cultured on uncoated porous titanium surfaces. Histologic observation of the implants showed that after 4 weeks healing, the bone to implant contact (BIC) on the LDc coated surface was much lower than that on the control surface, but didn't reach significant. In contrast, the percentage of bone within the implant's threads was significantly higher than the control group (P = 0.047). CONCLUSION: The BMP-2 gene coated sandblasted dual acid etched titanium implants slightly accelerated early bone formation around implants.

[Measurement of facial bone wall thickness of maxillary anterior teeth and premolars on cone beam computed tomography images].

OBJECTIVE: To measure the thickness of facial bone wall of maxillary anterior teeth and premolars based on cone beam computed tomography (CBCT) images. METHODS: CBCT images from 118 patients were collected from the Affiliated Stomatology Hospital of Zhejiang University. The thickness perpendicular to the long axis of facial bone wall was measured at two locations: 4 mm apical to the cementoenamel junction (point 1) and the middle of the root (point 2). RESULTS: The thickness of the facial bone walls of central incisors, lateral incisors and canines ranged from 0.5 to 1.5 mm. A thin facial bone wall (<1.0 mm) was quite frequent in central incisors (44.1% at point 1, 56.8% at point 2), lateral incisors (65.2% at point 1, 89.8% at point 2) and canines (45.8% at point 1, 61.0% at point 2). In contrast, the majority of examined first premolars (77.1% at point 1, 68.7% at point 2) and second premolars (94.1% at point 1, 94.1% at point 2) exhibited a thick facial bone wall (>1.0 mm). CONCLUSION: A thin facial bone wall of teeth in the anterior maxilla is common. Radiographic analysis of facial bone wall using CBCT is recommended for selection of appropriate treatment approach.

Osteogenesis of rat mesenchymal stem cells and osteoblastic cells on strontium-doped nanohydroxyapatite-coated titanium surfaces.

PURPOSE: The aim of this study was to compare the promotion of osteogenesis in vitro on three types of titanium surfaces: a strontium-hydroxyapatite (Sr-HA)-coated surface, a nano-HA-coated surface, and an uncoated roughened surface. MATERIALS AND METHODS: Sr-HA and HA were placed on disks with a roughened titanium surface by electrochemical deposition. MC3T3-E1 preosteoblast cells and rat bone mesenchymal stem cells were cultured on the Sr-HA, HA-coated, and uncoated roughened disks, and cell adhesion, proliferation, viability, osteogenic differentiation, and formation of mineralized nodules were measured at various time points. RESULTS: The Sr-HA coating produced by a simple electrochemical deposition treatment evidently enhanced the attachment, spreading, alkaline phosphatase activity, and extracellular matrix calcium mineralization of mouse bone mesenchymal stem cells and MC3T3-E1 cells compared with an untreated roughened titanium surface and a nano-HA-coated surface. CONCLUSION: This study suggests that a Sr-doped nano-HA coating produced through electrochemical deposition improves the osteoconductivity of a microrough titanium surface.

Analysis of facial bone wall dimensions and sagittal root position in the maxillary esthetic zone: a retrospective study using cone beam computed tomography.

PURPOSE: To determine the thickness of the facial bone wall and the sagittal angulation between the long axis of the teeth and the long axis of the associated alveolar bone in the maxillary esthetic zone using cone beam computed tomography (CBCT). MATERIALS AND METHODS: A retrospective radiographic study of CBCT images was conducted in 300 patients. The distance between the cementoenamel junction (CEJ) and the facial bone crest, the thickness of the facial bone wall (4 mm apical to the CEJ and midroot), and the sagittal angle between the long axis of teeth and the long axis of the respective alveolar bone were measured. Descriptive statistics and frequency analyses were performed. RESULTS: The sample included 133 men and 167 women (mean age, 36.9 years; range, 18 to 60 years). The distance between the CEJ and the facial bone crest varied between 0.1 and 4 mm and showed an overall tendency to increase with age. Almost 80% of anterior teeth and 40% of premolars exhibited a thin facial bone wall (< 1 mm), and nearly 30% of sites had a bone wall thinner than 0.5 mm. The sagittal angle at approximately 80% of central incisor and second premolar sites was < 20 degrees, but the angle at more than 40% of canine sites was ≥ 30 degrees. There was a significant decrease in facial bone wall thickness from premolars to anterior teeth. CONCLUSION: The facial bone wall in most maxillary anterior teeth was very thin. There was a substantial sagittal angulation between the long axes of teeth and those of their respective alveolar bone in most esthetic zone positions. CBCT analyses of the facial bone wall and the sagittal angle are recommended to ensure the most appropriate dental implant treatment approach.

Bone formation at porous titanium implants coated with multiple layers of recombinant human bone morphogenetic protein-2 cDNA plasmid in the posterior mandible in dogs.

PURPOSE: The objective of this study was to evaluate peri-implant bone formation and osseointegration at titanium implants coated with multiple layers of recombinant human bone morphogenetic protein-2 (rhBMP-2) cDNA placed into canine mandibles. MATERIALS AND METHODS: The rhBMP-2 plasmid was assembled on sandblasted/dual-acid-etched pure titanium implant surfaces using layer-by-layer assembly. The rhBMP-2 plasmid-coated implants were placed into the edentulous posterior mandibles of 12 adult beagle dogs. Implants without the rhBMP-2 plasmid coating served as controls. Treatments were randomized between jaw quadrants, and four implants were placed in each mandibular quadrant. After 4, 8, and 12 weeks in situ, mandibles were retrieved and prepared for removal torque testing and histomorphometric evaluation. RESULTS: Eight layers of rhBMP-2 plasmid were assembled onto the implant surfaces. Histomorphometric analysis showed that, after 4 and 8 weeks of healing, the intrathread bone area (BA) was slightly higher for test implants (54.21% and 59.56%) than for control implants (38.48% and 54.98%), respectively, but no statistically significant differences were seen at any time points. Mechanical tests showed that the mean removal torque values of the rhBMP-2 cDNA-coated implants were greater than those of the control implants after 8 weeks of healing (91 Ncm versus 61 Ncm; P = .31). CONCLUSION: Coating implants with multiple layers of an rhBMP-2 plasmid did not promote peri-implant bone formation and osseointegration in this model.

Effects of zinc-substituted nano-hydroxyapatite coatings on bone integration with implant surfaces.

OBJECTIVE: The purpose of this study was to investigate the effects of a zinc-substituted nano-hydroxyapatite (Zn-HA) coating, applied by an electrochemical process, on implant osseointegraton in a rabbit model. METHODS: A Zn-HA coating or an HA coating was deposited using an electrochemical process. Surface morphology was examined using field-emission scanning electron microscopy. The crystal structure and chemical composition of the coatings were examined using an X-ray diffractometer (XRD) and Fourier transform infrared spectroscopy (FTIR). A total of 78 implants were inserted into femurs and tibias of rabbits. After two, four, and eight weeks, femurs and tibias were retrieved and prepared for histomorphometric evaluation and removal torque (RTQ) tests. RESULTS: Rod-like HA crystals appeared on both implant surfaces. The dimensions of the Zn-HA crystals seemed to be smaller than those of HA. XRD patterns showed that the peaks of both coatings matched well with standard HA patterns. FTIR spectra showed that both coatings consisted of HA crystals. The Zn-HA coating significantly improved the bone area within all threads after four and eight weeks (P<0.05), the bone to implant contact (BIC) at four weeks (P<0.05), and RTQ values after four and eight weeks (P<0.05). CONCLUSIONS: The study showed that an electrochemically deposited Zn-HA coating has potential for improving bone integration with an implant surface.

Fluorescence microscopic analysis of bone osseointegration of strontium-substituted hydroxyapatite implants.

The purpose of this study was to analyze the effect of strontium-substituted hydroxyapatite (Sr-HA) on bone osseointegration of the implants using fluorescence microscopy. We allocated 20 implants to two groups: Sr-HA group and HA group. Electrochemically deposited HA and Sr-HA coatings were applied onto the implants separately. All the implants were inserted into femur bone of rabbits. Oxytetracycline hydrochloride, alizarin-complexon, and calcein green were respectively administered 7, 28, and 46 d after the implantation. After eight weeks, femurs were retrieved and prepared for the fluorescence microscopy observation. We analyzed the bone mineral apposition rates (MARs), bone area ratios (BARs), and bone to implant contact (BIC) of the two groups. Fluorescence microscopic observation showed that all groups exhibited extensive early peri-implant bone formation. The MAR of the Sr-HA group was greater than that for pure HA from 7 to 28 d after implantation, but no significant difference was found at later stage. And the BIC showed difference at 7 and 28 d compared with pure HA. We concluded that Sr-HA coating can improve the bone osseointegration of the implant in the early stage compared with the HA coating.

Effect of strontium-substituted nanohydroxyapatite coating of porous implant surfaces on implant osseointegration in a rabbit model.

PURPOSE: This study investigated the effects of a strontium-substituted nanohydroxyapatite (Sr-HA) coating, deposited onto porous implant surfaces using an electrochemical process, on implant osseointegration in a rabbit model. MATERIALS AND METHODS: The surfaces were analyzed by field-emission scanning electron microscopy, x-ray diffractometry (XRD), Fourier transform infrared spectroscopy (FT-IR), a portable surface roughness tester, and inductively coupled plasma atomic emission spectroscopy (ICP-AES). Thirty implants (half HA-coated and half Sr-HA-coated) were inserted into femurs of 15 rabbits. After 2, 4, and 8 weeks, the femurs were retrieved and prepared for histomorphometric evaluation. RESULTS: Microscopic examination showed a surface topography of rodlike crystals on both surfaces. XRD and FT-IR showed that the phase of the deposits was HA. No differences were found in surface roughness between the two groups. ICP-AES showed that the Sr/(Ca+Sr) molar ratio of Sr-HA coating was 10.1 mol%. Histologic observation showed that new bone appeared on both surfaces after 2 weeks and became mature after 8 weeks. Histomorphometric analysis showed no differences between the two groups in bone-to-implant contact at 2 weeks or in bone area within all threads at 2 and 4 weeks. The Sr-HA coated group had significantly higher bone-to-implant contact at 4 and 8 weeks. Significant differences were also found in bone area at 8 weeks. CONCLUSION: The present study showed that this Sr-HA coating, deposited using an electrochemical process, has the potential to enhance implant osseointegration.

Influence of multilayer rhBMP-2 DNA coating on the proliferation and differentiation of MC3T3-E1 cells seeded on roughed titanium surface.

For bone morphogenetic protein (BMP) gene therapy to be a viable approach for enhancing implant osseointegration clinically, requires the development of efficient nonviral delivery vectors that can coat the implant. This study evaluated a multilayer cationic liposome-DNA complex (LDc) coating as a delivery vehicle for recombinant human BMP-2 (rhBMP-2). Multilayered coatings, comprising hyaluronic acid (HA) and LDc, were fabricated onto titanium using a layer-by-layer (LBL) assembly technique. Preosteoblastic MC3T3-E1 cells were cultured on the roughened titanium surfaces coated with multilayers of HA/LDc, or on uncoated or HA/liposome only surfaces as controls. The amount of rhBMP-2 secreted by the MC3T3-E1 cells and the effect of the various surfaces on cell viability, proliferation, alkaline phosphatase (ALP) activity, osteocalcin (OC) secretion, and calcium deposition were evaluated. Messenger RNA levels of OC, ALP, Runx2, and Osx were also investigated. The results demonstrated that rhBMP-2 protein secreted into culture medium at 3 days was significantly higher than control groups. MC3T3-E1 cells cultured on the HA/LDc coating displayed significantly higher ALP activity and OC secretion at 7 days and 14 days culture, respectively. MC3T3-E1 cells cultured on HA/LDc upregulated expression of the osteoblast differentiation markers, especially on days 12 for OC and on days 6 and 12 for ALP and Osx. In conclusion, MC3T3-E1 cell cultured on the multilayer HA/LDc coating surface can secret rhBMP-2 protein and the protein levels were effective in inducing early osteogenic differentiation.